Search

CN-121987590-A - Lipid nanoparticle comprising adjuvant for mucosal administration and method for preparing the same

CN121987590ACN 121987590 ACN121987590 ACN 121987590ACN-121987590-A

Abstract

The invention relates to lipid nano-particles containing an adjuvant for mucosal administration and a preparation method thereof. The lipid nanoparticle comprises cationic lipid C12-200, total lipid consisting of non-cationic phospholipid, cholesterol and derivatives thereof and polyethylene glycol lipid, and an adjuvant and an active ingredient, wherein the adjuvant is selected from at least one of AS03, MF59, monophosphoryl lipid A, microminiaturized monophosphoryl lipid A, ophiopogonin D, saponin QS21, QS7, QS17, QS18, cpG oligonucleotide, 2 '3' -cyclic GMP-AMP, 3 '3' -cyclic GMP-AMP, alpha GC, trehalose-6,6-dibehenate, cholera toxin B subunit and flagellin.

Inventors

  • SUN XIAO

Assignees

  • 珠海丽凡达生物技术有限公司

Dates

Publication Date
20260508
Application Date
20260202

Claims (10)

  1. 1. Lipid nanoparticle comprising an adjuvant for mucosal administration, characterized in that the lipid nanoparticle comprises a total lipid consisting of cationic lipids C12-200, non-cationic phospholipids, cholesterol and derivatives thereof and pegylated lipids, and an adjuvant and an active ingredient; Wherein the adjuvant is selected from at least one of AS03, MF59, monophosphoryl lipid A, miniaturized monophosphoryl lipid A, ophiopogonin D, saponin QS21, QS7, QS17, QS18, cpG oligonucleotide, 2 '3' -cyclic GMP-AMP, 3 '3' -cyclic GMP-AMP, αGC, trehalose-6,6-dibehenate, cholera toxin B subunit, flagellin; the non-cationic phospholipids include anionic phospholipids and neutral phospholipids; the active ingredient is a molecule with therapeutic, prophylactic or diagnostic functions, including but not limited to nucleic acid drugs, small molecule drugs, polypeptide/protein drugs, gene editing tools.
  2. 2. The lipid nanoparticle of claim 1, wherein the anionic phospholipid is selected from the group consisting of DOPS, 1, 2-dioleoyl-sn-glycero-3-phospho-L-serine, 1, 2-di-arachidonoyl-sn-glycero-3-phospho-L-serine, 1, 2-dioleoyl-sn-glycero-3-phospho-L-homoserine, 1, 2-dioleoyl-sn-glycero-3-phospho-O-methyl-L-serine, 1, 2-dioleoyl-sn-glycero-3-phospho-O-phospho-L-homoserine, and the neutral phospholipid is selected from the group consisting of phosphatidylcholine and derivatives thereof, phosphatidylethanolamine and derivatives thereof, sphingomyelin; Preferably, the phosphatidylcholine and the derivatives thereof are selected from DMPC, DPPC, DSPC, DAPC, DBPC, POPC, DOPC, SOPC, DLPC, preferably, the phosphatidylethanolamine and the derivatives thereof are selected from DOPE, DPPE, POPE, preferably, the sphingomyelin is selected from PSM and cephalin; Preferably, the anionic phospholipid is DOPS, and preferably, the neutral phospholipid is DSPC.
  3. 3. The lipid nanoparticle of claim 1, wherein the cationic lipid comprises 20 mol% -49.5% mol% of the total lipid, the non-cationic phospholipid comprises 13% mol% -30% mol% of the total lipid, the cholesterol and derivatives thereof comprise 15.5% mol% -65% mol% of the total lipid, and the pegylated lipid comprises 2% mol% -5% mol% of the total lipid; preferably, the anionic phospholipid accounts for 8 mol-30 mol% of the total lipid, and the neutral phospholipid accounts for not more than 50 mol% of the non-cationic lipid; Preferably, the anionic phospholipid comprises 8 mol% of the total lipid and the neutral phospholipid comprises 8 mol% of the total lipid.
  4. 4. The lipid nanoparticle of claim 1, wherein the adjuvant is at least one of αgc, cpG, saponin QS 21; Preferably, the adjuvant is CpG, and preferably, the mass ratio of the CpG to the active ingredient is 1:4.
  5. 5. A method for preparing lipid nanoparticles comprising an adjuvant for mucosal administration according to claims 1-4, comprising (1) dissolving anionic phospholipids in solvent a to obtain lipid solution 1, dissolving other lipids in anhydrous C1-C4 lower alcohols to obtain lipid solution 2, adding lipid solution 2 to lipid solution 1 to obtain lipid working fluid; (2) The active ingredient is RNA, and the RNA and CpG are dissolved in citrate buffer solution or sodium acetate buffer solution to obtain an active ingredient solution; (3) Uniformly mixing the active ingredient solution and the lipid working solution to obtain a mixed solution, and removing anhydrous C1-C4 low-carbon alcohol in the lipid working solution to obtain the lipid nanoparticle containing the adjuvant for mucosal administration; solvent A is selected from enzyme-free water, DMSO, N-methyl-2-pyrrolidone, propylene glycol, and polyethylene glycol 400; Preferably, the solvent a is selected from enzyme-free water.
  6. 6. A method of preparing lipid nanoparticles comprising an adjuvant for mucosal administration according to claim 5, wherein the concentration of mRNA in the active ingredient solution is 80 μg/mL and the concentration of CpG is 20 μg/mL.
  7. 7. The method for preparing lipid nanoparticles comprising an adjuvant for mucosal administration according to claim 6, wherein the mixing ratio of the active ingredient solution and the lipid working solution in the step (3) is (2-4): 1 (v/v).
  8. 8. A method of preparing lipid nanoparticles comprising an adjuvant for mucosal delivery according to claim 6, wherein in step (3), the mixing of the active ingredient solution and the lipid working fluid is preferably performed by microfluidic device, and the flow rate is controlled to be 6-24 mL/min, more preferably 12mL/min.
  9. 9. An adjuvant-containing lipid nanoparticle composition for mucosal administration, comprising the adjuvant-containing lipid nanoparticle for mucosal administration prepared according to claims 1 to 4 or according to claims 5 to 8, or a pharmaceutically acceptable carrier, diluent, excipient or propellant; preferably, the lipid nanoparticle composition comprising an adjuvant for mucosal administration is for nasal mucosal administration, oral mucosal administration, sublingual mucosal administration, ocular mucosal administration, rectal mucosal administration, vaginal mucosal administration and any combination thereof; preferably, the lipid nanoparticle composition for mucosal administration comprising an adjuvant is for nasal mucosal administration, the composition being formulated as a nasal gel, nasal spray, nasal drops, inhalation aerosol, inhalation powder mist or an aerosol inhalation solution; Preferably, the lipid nanoparticle composition for mucosal administration comprising an adjuvant is for buccal or sublingual mucosal administration, said composition being formulated as an oral film, sublingual tablet, buccal tablet or oral spray; preferably, the lipid nanoparticle composition comprising an adjuvant for mucosal administration is used for preventing respiratory viral infections, for treating respiratory diseases or for treating local respiratory infections.
  10. 10. Use of a lipid nanoparticle composition comprising an adjuvant for mucosal administration according to claim 9 for the manufacture of a medicament for the prevention and/or treatment of a disease or disorder caused by any one or more of the following causes: (i) Infectious diseases, local or systemic infections caused by viruses, bacteria, fungi or parasites; (ii) Central nervous system diseases such as neurodegenerative diseases, epilepsy, brain tumor, apoplexy, mental diseases or central nervous system injury; (iii) Inflammatory and immune diseases, autoimmune diseases, allergic diseases, asthma, inflammatory bowel diseases or arthritis; (iv) Metabolic diseases such as diabetes, obesity or dyslipidemia; (v) Neoplastic diseases, immunotherapy, gene therapy or chemotherapy for solid or hematological tumors; (vi) Genetic diseases, which are monogenic genetic diseases or diseases caused by abnormal functions of specific genes; (vii) Pain, acute pain, chronic pain or neuropathic pain.

Description

Lipid nanoparticle comprising adjuvant for mucosal administration and method for preparing the same Technical Field The technical field of the invention is the technical field of medicine. In particular to lipid nano-particles containing an adjuvant for mucosal administration and a preparation method thereof. Background Mucosal immunity is used as a first defense line for preventing pathogen invasion of organisms, and covers key portals such as respiratory tract, digestive tract, genitourinary tract and the like. However, existing vaccines (including subunit vaccines and novel platforms such as nucleic acid vaccines) are mainly inoculated by intramuscular or subcutaneous injection, the immune response induced by the existing vaccines is mainly systemic, and efficient specific immune response is difficult to be induced locally in mucous membranes, and particularly the level of secretory immunoglobulin A (sIgA) is obviously insufficient. This results in the vaccine failing to establish an effective immune barrier at the initial site of pathogen invasion, thereby limiting the protective effect against mucosal transmitted pathogens such as influenza virus, SARS-CoV-2, norovirus and HIV. Lipid nanoparticles have shown good encapsulation and delivery capacity in systemic administration as important delivery vehicles for nucleic acid based vaccines. However, if the composition is directly applied to a mucosal administration scene, challenges such as low delivery efficiency, insufficient immune activation and the like are often faced, and an ideal mucosal immune effect is difficult to realize. In mucosal vaccine design, the choice and compatibility of the adjuvant is particularly important. The ideal mucosal adjuvant needs to have multiple functions at the same time, namely overcoming the local immune tolerance state of the mucous membrane, effectively activating antigen presenting cells such as dendritic cells and the like, and finely regulating the direction of immune response so as to lead the antigen presenting cells to tend to generate Th2 or Th17 type immune response mainly comprising sIgA. However, the integration of the adjuvant with the delivery vehicle is not a simple physical mixing process. There is a complex interaction between the physicochemical properties, mechanism of action, and the composition, structure, and surface properties of the lipid nanoparticle. These interactions directly affect the loading efficiency of the adjuvant in the carrier, the release kinetics, and ultimately the spatiotemporal delivery synergy with the antigen. Different adjuvants may activate immune cells through different pattern recognition receptors, with different sites of action (intracellular or membrane) and desired intracellular delivery efficiencies. Thus, not any adjuvant in combination with lipid nanoparticles can produce synergistically enhanced immune effects. Disclosure of Invention In view of the above, it is an object of the present invention to provide an adjuvant-containing lipid nanoparticle for mucosal administration, and it is a second object of the present invention to provide a method for preparing the adjuvant-containing lipid nanoparticle for mucosal administration. In order to achieve the above purpose, the present invention provides the following technical solutions: Lipid nanoparticles comprising an adjuvant for mucosal administration comprising a total lipid consisting of cationic lipids C12-200, non-cationic phospholipids, cholesterol and derivatives thereof and pegylated lipids, an adjuvant and an active ingredient, wherein the adjuvant is selected from the group consisting of AS03, MF59, monophosphoryl lipid a, miniaturized monophosphoryl lipid a, ophiopogonin D, saponin QS21, QS7, QS17, QS18, cpG oligonucleotides, 2 '3' -cyclic GMP-AMP, 3 '3' -cyclic GMP-AMP, αgc, trehalose-6,6-dibehenate, cholera toxin B subunit, flagellin, and at least one of anionic phospholipids and neutral phospholipids. In a preferred embodiment, the anionic phospholipids are selected from the group consisting of 1, 2-dioleoyl-sn-glycero-3-phosphate-L-serine (DOPS), 1, 2-dioleoyl-sn-glycero-3-phosphate-L-serine, 1, 2-dioleoyl-sn-glycero-3-phosphate-L-homoserine, 1, 2-dioleoyl-sn-glycero-3-phosphate-O-methyl-L-serine, 1, 2-dioleoyl-sn-glycero-3-phosphate-O-phosphate-L-homoserine, and the neutral phospholipids are selected from the group consisting of phosphatidylcholine and derivatives thereof, phosphatidylethanolamine and derivatives thereof, sphingomyelin. In a preferred embodiment, the phosphatidylcholine and its derivatives are selected from DMPC, DPPC, DSPC, DAPC, DBPC, POPC, DOPC, SOPC, DLPC. In a preferred embodiment, the phosphatidylethanolamine and derivatives thereof are selected from DOPE, DPPE, POPE. In a preferred embodiment, the sphingomyelin is selected from PSM, cephalin. In a preferred embodiment, the anionic phospholipid is DOPS. In a preferred embodiment, the neutral phospholipid is DSPC. In a preferre